Figure 7
Expression levels of the metalloprotease responsible for cleavage of Ebola glycoprotein. Shown are (a) expression levels of tumor necrosis factor (TNF)-α converting enzyme/α-disintegrin and metalloproteinase (ADAM)-17 from the overview cluster and (b) in graph form. (c) Glycoprotein (GP) in the serum from infected rhesus macaques over the course of infection. Serum was diluted 1:3 in NP40 lysis buffer. Samples were run on a 10% Bis-Tris gel under reducing conditions, as shown. Mock cell lysate from 293T cells transfected with vector only (pDisplay) is shown as a negative control (lane 1); Zaire Ebola virus (ZEBOV; lane 2) is supernatant from in vitro Ebola infected Vero E6 cells at day 8 after infection. Lanes 3 and 4 are transfection controls expressing glycoprotein (GP)1,2 (cell lysate) and GP1,2Δ supernatant) [31]. Serum from infected rhesus macaques, before infection (lane 5), and on day 4 and 6 after infection (lanes 6 to 9) were diluted 1:3 in NP40 lysis buffer and 22.5 μl was loaded per lane. Samples included two animals per day (after infection) analyzed. Note the lack of GP in the prebleed control sample (lane 5). GP2Δ is seen in the transfection control (lane 4) and NHP sera samples from days 4 (lane 6, albeit weakly) and 6 days after infection (lanes 8 and 9).